Effects of Hydrogen Sulfide on Inflammatory Factors and Mitochondrial Energy Metabolic Disorders After Reperfusion Injury in Rats

Abstract

Objective To explore the effect of hydrogen sulfide on inflammatory factors and energy metabolism of mitochondria after limbs reperfusion injury in rats. Methods Sixty rats were divided into three groups:sham operation group,control group(ischemia-reperfusion injury + saline group),and experimental group(ischemia-reperfusion injury + H2S group).Wistar rat models of limb ischemia-reperfusion injury were established.Skeletal muscle samples were collected to determine the levels of necrosis decomposition products [including myoglobin(MB),lipoprotein complex(LPC)and lipid peroxide(LPO)];blood samples were collected to determine the levels of interleukin(IL)-1,IL-6 and tumor necrosis factor-α(TNF-α);mitochondria were extracted for mitochondrial transmembrane potential measurement and ATP content detection.Statistical analysis was made on the test results. Results After ischemia reperfusion injury,the levels of MB,LPO,and LPC in skeletal muscle,liver,lung and renal tissues of the control group were significantly increased(MB:Pskeletal muscle =0.003,Pliver =0.001,Plung =0.001,Pkidney =0.001;LPO:Pskeletal muscle =0.001,Pliver =0.001,Plung =0.001,Pkidney =0.002;LPC:Pskeletal muscle =0.000,Pliver =0.002,Plung =0.002,Pkidney =0.003),and hydrogen sulfide treatment during ischemia reperfusion significantly inhibited the production of MB,LPO,and LPC(MB:Pskeletal muscle =0.021,Pliver =0.036,Plung =0.005;LPO:Pskeletal muscle =0.003,Pliver =0.008,Plung =0.010,Pkidney =0.015;LPC:Pskeletal muscle =0.002,Pliver =0.026,P lung =0.007,P kidney =0.006).Ischemia/reperfusion of lower extremity in rats resulted in increased levels of IL-1,IL-6,and TNF-α in the serum of rats,and the levels of IL-1,IL-6,and TNF-increased over time,with statistically significant differences in IL-1,IL-6,and TNF-α among groups at 3 h(IL-1:P3 h =0.019,P6 h =0.011,P9 h =0.009,$P_{12_{h}}$=0.008,and P15 h =0.002;IL-6:P3 h =0.026,P6 h =0.009,P9 h =0.002, $P_{12_{h}}$=0.002,P15 h =0.003;TNF-α:P3 h =0.002,P6 h =0.002,P9 h =0.005,$P_{12_{h}}$=0.002,P15 h =0.003).The levels of IL-1,IL-6,and TNF-α in serum were significantly inhibited during ischemia reperfusion(IL-1:P3 h =0.035,P6 h =0.039,P9 h =0.012,$P_{12_{h}}$=0.005,P15 h =0.006;IL-6:P3 h =0.042,P6 h =0.025,P9 h =0.023,$P_{12_{h}}$=0.006,P15 h =0.005;TNF-α:P3 h =0.005,P6 h =0.003,P9 h =0.022,$P_{12_{h}}$=0.005,P15 h =0.005),and such inhibitory effects became even more obvious over time.After limb ischemia and reperfusion in the control group,the mitochondrial transmembrane potential of skeletal muscle cells significantly decreased compared with that of the sham group(t=6.698;P=0.001).After hydrogen sulfide treatment,the mitochondrial membrane potential energy of the experimental group was significantly higher than that of the control group(t=7.507,P = 0.000).The ATP level in the mitochondria of ischemia reperfusion rats in the control group was significantly lower than that in the sham group(t=7.526,P = 0.000).The content of mitochondrial ATP in the experimental group was significantly higher than that in the control group after hydrogen sulfide treatment(t=8.604,P = 0.000). Conclusions Hydrogen sulfide can alleviate the injury of skeletal muscle and distal organs after limb ischemia-reperfusion and reduce local inflammatory reaction.In addition,it is valuable in alleviating mitochondrial transmembrane potential and energy metabolism disorders during reperfusion injury.